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Conformational changes associated to sensing mechanisms of heme-based protein sensors are a key molecular event that seems to modulate not only the protein activity but also the potential of the Fe III/II redox couple of the heme domain. In this work, midpoint potentials ( E m ) assigned to the Fe III/II redox couple of the heme domain of FixL from Rhizobium etli ( Re FixL) in the unliganded and liganded states were determined by spectroelectrochemistry in the presence of inorganic mediators. In comparison to the unliganded Re FixL protein (+ 19 mV), the binding to ligands that switch off the kinase activity induces a negative shift, i. e. E m = − 51, − 57 and − 156 mV for O 2 , imidazole and CN − , respectively. Upon binding to CO, which does not affect the kinase active, E m was observed at + 21 mV. The potential values observed for Fe III/II of the heme domain of Re FixL upon binding to CO and O 2 do not follow the expected trend based on thermodynamics, assuming that positive potential shift would be expected for ligands that bind to and therefore stabilize the Fe II state. Our results suggest that the conformational changes that switch off kinase activity upon O 2 binding have knock-on effects to the local environment of the heme, such as solvent rearrangement, destabilize the Fe II state and counterbalances the Fe II -stabilizing influence of the O 2 ligand. [ABSTRACT FROM AUTHOR]

Abstract: Brassinosteroids (BRs) are steroid hormones that participate in multiple biological processes. In this paper, we characterized a classic rice mutant Fn189 (dwarf54, d54) showing semi-dwarf stature and erect leaves. The coleoptile elongation and root growth was less affected in Fn189 than wild-type plant by the exogenous application of eBL, the most active form of BRs. Lamina joint inclination assay and morphological analysis in darkness further showed that Fn189 mutant plant was insensitive to exogenous eBL. Through map-based cloning, Fn189 was found to be a novel allelic mutant of the DWARF 61 (D61) gene, which encodes the putative BRs receptor OsBRI1. A single base mutation caused the I834F substitution in the OsBRI1 kinase domain. Consequently, kinase activity of OsBRI1 was found to decrease dramatically. Taken together, the kinase activity of OsBRI1 is essential for brassinosteroids to regulate normal plant growth and development in rice. [Copyright &y& Elsevier]